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• W1580095901 abstract "Abstract One of the most fundamental concepts of microscopy is that of resolution–the ability to clearly distinguish two objects as separate. Recent advances such as structured illumination microscopy (SIM) and point localization techniques including photoactivated localization microscopy (PALM), and stochastic optical reconstruction microscopy (STORM) strive to overcome the inherent limits of resolution of the modern light microscope. These techniques, however, are not always feasible or optimal for live cell imaging. Thus, in this review, we explore three techniques for extracting high resolution data from images acquired on a widefield microscope–deconvolution, model convolution, and Gaussian fitting. Deconvolution is a powerful tool for restoring a blurred image using knowledge of the point spread function (PSF) describing the blurring of light by the microscope, although care must be taken to ensure accuracy of subsequent quantitative analysis. The process of model convolution also requires knowledge of the PSF to blur a simulated image which can then be compared to the experimentally acquired data to reach conclusions regarding its geometry and fluorophore distribution. Gaussian fitting is the basis for point localization microscopy, and can also be applied to tracking spot motion over time or measuring spot shape and size. All together, these three methods serve as powerful tools for high‐resolution imaging using widefield microscopy. J. Cell. Physiol. 229: 132–138, 2014. © 2013 Wiley Periodicals, Inc." @default.
• W1580095901 created "2016-06-24" @default.
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• W1580095901 date "2013-10-25" @default.
• W1580095901 modified "2023-10-16" @default.
• W1580095901 title "Bending the Rules: Widefield Microscopy and the Abbe Limit of Resolution" @default.
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• W1580095901 doi "https://doi.org/10.1002/jcp.24439" @default.
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